硫代磷酸寡脱氧核苷酸体外抗乙型肝炎病毒的实验研究

Inhibitory effect of phosphorothioate oligodeoxynucleotides on HBV replication and synthesis of antigen in vitro

目的了解三螺旋形成寡核苷酸（TFO）、反义寡核苷酸（ODNas）抗乙型肝炎病毒（HBV）作用。方法设计合成针对HBV核心启动子1734nt-1754nt及前C、前基因组RNA5’端起始区的21聚硫代磷酸TFD（TFO21）及21聚硫代磷酸ODNas（ODNas21）。在22．1．5细胞观察了各寡核苷酸的抗HBV作用。结果TFO21、ODNas21处理的22．1．5细胞HBsAg、HBeAg及HBVDNA分泌明显低于对照组。浓度为10μmol／L的TFO21与ODNas21对HBsAg、HBeAg的抑制分别达57．5％、77．0％；61．0％、79．6％。两者联合给药对HBsAg、HBeAg的抑制分别达71．5％、85．0％。该抑制呈剂量依赖性。无关序列对照寡脱氧核苷酸光明显抑制作用。在实验范围内，硫代寡核苷酸对22．1．5细胞无毒性作用。结论在HBV细胞模型系统，TFO21、ODNas21能有效抑制HBV复制及抗原合成，具有较大应用潜力。

Objective To evaluate the effect of triplex forming oligodeoxyncleotides (TFO) and antisense oligodeoxynucleotides (ODNas) on the replication of HBV. Methods A 21mer phosphorothioate TFO (TFO21)directed at 1734nt-1754nt sites in HBV core promoter and a 21 mer phosphorothioate ODNas (ODNas21 )complementory to the initiation sites of pre C RNA and pregenomic RNA were synthesized. Effect of TFO21 and ODNas21 on HBV replication and synthesis of antigen were observed in 22.1.5 cells. Results Both TFO21 and ODNas21 showed the inhibition to HBV replication and synthesis of antigen while, ODNcon (control of 21 mer phosphorothioate oligodeoxynucleotieds ) showed little inhibitory effects. At concentration of 10 μmol/L, TFO21and ODNas21 inhibited the synthesis of HBsAg and HBeAg by 57.5% and 77%; 61% and 79 .6%, respectively.The mixture of TFO21 and ODNas21 was more effective than TFO21 or ODNas21 alone. The inhibitions were dosedependent. No toxicity was observed in the 22. 1 .5 cells treated with those oligodeoxyncleotides. Conclusion Triplex forming oligodeoxyncleotides and antisense oligodeoxyncleotides were both potent inhibitor, for HBV replication and synthesis of antigen.